Characterization of a monoclonal antibody recognizing the E1α subunit of plant mitochondrial pyruvate dehydrogenase

Michael H. Luethy, Nancy R. David, Thomas E. Elthon, Jan A. Miernyk, Douglas D. Randall

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

We have isolated a monoclonal antibody that recognizes the E1α subunit of the plant mitochondrial pyruvate dehydrogenase complex. The antibody specifically recognizes the Ela subunit from maize seedling, pea leaf, and castor oil seed endosperm mitochondrial pyruvate dehydrogenases, but does not recognize the E1α subunit present in the plastid complexes from these plants. The pea mitochondrial pyruvate dehydrogenase complex was used for subsequent characterization of the antibody. Two-dimensional electrophoretic analysis of a phosphorylated pea mitochondrial pyruvate dehydrogenase complex preparation revealed that the monoclonal antibody recognizes all phosphorylated forms of the E1α subunit. Under these conditions, the only proteins recognized by the antibody are phosphorylated. Binding of the antibody to the pyruvate dehydrogenase complex inhibits both catalytic activity and phosphorylation of the E1α subunit, but does not significantly inhibit dephosphorylation. The monoclonal antibody recognizes mitochondrial E1α subunits from a variety of plant materials including monocot and dicot seedlings, and thermogenic and storage tissues. The antibody does not recognize the E1α subunit from rat liver or pig heart mitochondria, yeast, or bacteria. This highly specific antibody will be a useful tool for study of plant mitochondrial pyruvate dehydrogenase complexes.

Original languageEnglish (US)
Pages (from-to)443-449
Number of pages7
JournalJournal of Plant Physiology
Volume145
Issue number4
DOIs
StatePublished - Jan 1 1995

Fingerprint

pyruvate dehydrogenase (lipoamide)
Pyruvate Dehydrogenase Complex
Pyruvic Acid
monoclonal antibodies
Oxidoreductases
Monoclonal Antibodies
antibodies
Antibodies
Peas
peas
Seedlings
Castor Oil
castor oil
Heart Mitochondria
Endosperm
Plastids
leaf oils
seedlings
dephosphorylation
Magnoliopsida

Keywords

  • MAb
  • Mitochondria
  • PDC
  • PDH
  • Pisum sativum L.
  • ZmPDHα
  • monoclonal antibody
  • monoclonal antibody
  • monoclonal antibody generated using the maize PDH alpha subunit as the antigen
  • protein pbospborylation
  • pyruvate debydrogenase complex
  • pyruvate dehydrogenase
  • pyruvate dehydrogenase complex

ASJC Scopus subject areas

  • Physiology
  • Agronomy and Crop Science
  • Plant Science

Cite this

Characterization of a monoclonal antibody recognizing the E1α subunit of plant mitochondrial pyruvate dehydrogenase. / Luethy, Michael H.; David, Nancy R.; Elthon, Thomas E.; Miernyk, Jan A.; Randall, Douglas D.

In: Journal of Plant Physiology, Vol. 145, No. 4, 01.01.1995, p. 443-449.

Research output: Contribution to journalArticle

Luethy, Michael H. ; David, Nancy R. ; Elthon, Thomas E. ; Miernyk, Jan A. ; Randall, Douglas D. / Characterization of a monoclonal antibody recognizing the E1α subunit of plant mitochondrial pyruvate dehydrogenase. In: Journal of Plant Physiology. 1995 ; Vol. 145, No. 4. pp. 443-449.
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abstract = "We have isolated a monoclonal antibody that recognizes the E1α subunit of the plant mitochondrial pyruvate dehydrogenase complex. The antibody specifically recognizes the Ela subunit from maize seedling, pea leaf, and castor oil seed endosperm mitochondrial pyruvate dehydrogenases, but does not recognize the E1α subunit present in the plastid complexes from these plants. The pea mitochondrial pyruvate dehydrogenase complex was used for subsequent characterization of the antibody. Two-dimensional electrophoretic analysis of a phosphorylated pea mitochondrial pyruvate dehydrogenase complex preparation revealed that the monoclonal antibody recognizes all phosphorylated forms of the E1α subunit. Under these conditions, the only proteins recognized by the antibody are phosphorylated. Binding of the antibody to the pyruvate dehydrogenase complex inhibits both catalytic activity and phosphorylation of the E1α subunit, but does not significantly inhibit dephosphorylation. The monoclonal antibody recognizes mitochondrial E1α subunits from a variety of plant materials including monocot and dicot seedlings, and thermogenic and storage tissues. The antibody does not recognize the E1α subunit from rat liver or pig heart mitochondria, yeast, or bacteria. This highly specific antibody will be a useful tool for study of plant mitochondrial pyruvate dehydrogenase complexes.",
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